Study On The Preparation And Chemical Constituents Of Anti-livestock And Poultry Mycoplasma Of Traditional Chinese Medicine Compound Fei Kangming Granule

Mycoplasma caused by pigs mycoplasma pneumonia, chicken poison mycoplasma disease, infectious bovine pleuropneumonia and goat contagious pleuropneumonia breeding to bring serious economic loss. Due to the inefficiency of the current mycoplasma vaccine immunization, tylosin of veterinary clinical commonly used antimicrobial drug resistance and drug residues serious problems, which were widely attention. It need to develop cheap, effective, broad-spectrum, non-toxic anti-livestock and poultry mycoplasma of traditional Chinese medicine compound preparation urgently. For the further development of Mycoplasma animal compound in this subject in the preliminary work on the basis of selected South medicine Zanthoxyhim nitidum (Roxb.) DC, Pogostemon cablin (Blanco) Benth., Baphicacanthus cusia (Nees) Bremek., Coptis chinensis Franch., Scutellaria baicalensis Georgi., Cortex Phellodendri Chinensis, Glycyrrhiza Fisch., Dictamnus dasycarpus Turcz., Pseudolarix kaempferi Gord. and Artemisia argyi Levl. et Vant,10kinds of traditional Chinese medicine such as artemsia argyi. By micro drug sensitivity test in vitro, with veterinary rationale and pharmaceutics of TCM theory, screening effective resistance to livestock and poultry mycoplasma single taste traditional Chinese medicine and the best of traditional Chinese medicine compound. Through the pharmacodynamics test to evaluate the safety and clinical efficacy of the best traditional Chinese medicine compound, and made into traditional Chinese medicine compound granule, and studied on quality standard, preliminary stability and chemical constituents for traditional Chinese medicine compound granule. Specific research methods and results were as follows.1Anti-livestock and poultry mycoplasma of traditional Chinese medicine compound Fei kangming established Using water refluxing extraction and the micro-liquid dilution method, researched on Zanthoxylum nitidum (Roxb.) DC, Pogostemon cablin (Blanco) Benth., Baphicacanthus cusia (Nees) Bremek., Coptis chinensis Franch., Scutellaria baicalensis Georgi., Cortex Phellodendri Chinensis, Glycyrrhiza Fisch., Dictamnus dasycarpus Turcz., Pseudolarix kaempferi Gord. and Artemisia argvi Levl. et Vant., folium artemisiae argyi10kinds such as traditional Chinese medicine for anti-livestock and poultry mycoplasma in vitro. Results showed that10of traditional Chinese medicine, with Pogostemon eablin (Blanco) Benth., Pseudolarix kaempferi Gord. and Artemisia argyi Levl. et Vant., the remaining7taste traditional Chinese medicine of three kinds of livestock and poultry mycoplasma had varying degrees of restrain and kill effect in vitro; Controlled drugs tartaric acid tylosin to experiment with three kinds of livestock and poultry mycoplasma had higher sensitivity. For effective resistance to livestock and poultry mycoplasma single taste traditional Chinese medicine two joint drug sensitivity test, the results showed that7flavour combination and compatibility of Chinese water extraction liquid two of cattle mycoplasma, chicken poison mycoplasma and mycoplasma pneumoniae the inhibition effect of sheep shows Zanthoxylum nitidum (Roxb.) DC, Baphic acanthus cusia (Nees) Bremek., Coptis chinensis Franch., ScuteUaria baicalensis Georgi., Cortex Phellodendri Chinensis, Glycyrrhiza Fisch. and Dictamnus dasycarpus Turcz., no antagonism between appropriate compatibility. Under the guidance of TCM theory, according to the prescriptions of traditional Chinese medicine composition, principle and the compatibility of the law from7groups of compound Chinese medicine, through the study drug sensitive test showed that the compound Chinese medicine compound and7group were with different degrees of livestock and poultry mycoplasma resistance activities, including compound5(Baphicacanthus cusia (Nees) Bremek., Zanthoxylum nitidum (Roxb.) DC, Dictamnus dasycarpus Turcz.) had best effect, of which the ingredients were less, so choosed compound5as best anti-livestock and poultry mycoplasma of traditional Chinese medicine compound, named compound Fei kangming.2Sudy on pharmacodynamics of traditional Chinese medicine compound Fei kangmingHealthy chicken for clinical drug safety studies, results show that traditional Chinese medicine compound Fei kangming to16g/d water drinking5d, visceral organs of chickens without any exception, prompting the party this dose on chicken safety. It adopted1×109MG of CCU/mL chicken breasts balloon inoculation of0.5mL, nasal drops0.3mL, point the eyes0.2ml., and created stress conditions to build artificial poison mycoplasma infection chicken chicken model, and through the clinical symptoms and pathological observation, isolation, culture, identification and serological diagnosis, confirmed the disease models were successful. Clinical efficacy of the test results showed that the compound Fei kangming high, middle dose group of MG artificial infection chicken with obvious therapeutic effect. Mortality compared with infection control group, the treatment group had varying degrees of decline; Compound Fei kangming high, middle dose group and controlled drugs tartaric acid tylosin group were0%. Compound Fei kangming low dose group and prevention group mortality rate were very low, which were5.00%and2.50%respectively. The cure rate, compound Fei kangming curative effect of high dose group (92.5%), the best were better than low dose group, the dose group and tartaric acid tylosin controlled drugs had the same effect (90%). Weight gain, the treatment group were significantly higher than control group of infection, but lower than that of normal control group, compound Fei kangming high-dose group and normal control group; Feed conversion rate, tartaric acid tylosin group were lowest, the other drug group and health groups quitily, infection control group were highest.Serum antibody levels, the treatment group can significantly reduced the antibody positive reaction, which the best compound Fei kangming high-dose group. Balloon injury, each treatment group airbag lesions were lighter than infection control group, which the best compound Fei kangming high-dose group, followed by compound Fei kangming middle-dose group and tartaric acid tylosin group. This compound for MG infection had good treatment effect, and to overcome the shortcoming of antibiotic drug using for a long time which will produce resistance, which could be used in the prevention and cure of the disease. Recommend compound Fei kangming0.4%for the treatment of MG chickens against the water drinking to drug concentration,0.2%prevention against the drinking water to drug concentration, continuous administration5d, with every day in a liquid.3Study on preparation of granules of traditional Chinese medicine compound Fei kangmingAccording to clinical need, the results of the preliminary experiments, production conditions and technical level, choosed granule as dosage form. This experiment was mainly to multi-index comprehensive evaluation more ingredients, orthogonal experiment design and single factor investigation method for screening the best preparation process, the final preparation of traditional Chinese medicine compound Fei kangming particles were as follows, took prescription proportion (Baphicacanthus cusia (Nees) Bremek., Zanthoxylum nitidum (Roxb.) DC, Dictamnus dasycarpus Turcz.(10mesh), precision said, adding12times the amount of water, soaking0.5h, reflux extracting3times, each time1.5h, merge three extracts, the filtrate overspeed centrifugal (8000r/min,15min), vacuum decompression (temperature60℃vacuum degree was0.08～0.10MPa, steam pressure0.10MPa) or less concentrated to the relative density of1.10(60℃) liquid extract; Took the liquid extract for spray drying, the material temperature:50～60℃, liquid speed:50-60mL/min, inlet air temperature:165～170℃, the outlet air temperature:70～75℃, collected dry extract powder. According to the dry extract powder, dextrin1:0.6blending, through80mesh sieve, the dry granulating method was adopted, set roll pressure was60～70kgf/’cm2, roll speed was12～14r/min, powder feeding speed was14～16r/min. It made from granule1200g, sieving, packaging, quick. Meantime proved by three batch of pilot trial production, the proposed was simple and reasonable, stable and practical, could meet the requirements of industrial production.4Study on quality standard and preliminary stability of traditional Chinese medicine compound Fei kangming granulesQuality standard research, the main raw material medicine and complementary makings quality standards and granules quality standards research. Raw medicinal materials and auxiliary materials mainly in the quality standard for feeding medicine yinpian in accordance with the law of the People’s Republic of China pharmacopoeia2010edition (1) under the relevant provisions of inspection. Accessories-dextrin in accordance with the law of the People’s Republic of China pharmacopoeia2010edition (2) under the relevant provisions of inspection. Granules in the quality standard of pilot production of three batch samples for identification, examination and content determination of research. Each other all the herbs in the TLC identification test, and let Baphicacanthus cusia (Nees) Bremek., Zanthoxylum nitidum (Roxb.) DC, Dictamnus dasycarpus Turcz. of TLC identification on the body of this product quality standard. Under inspection, on the particle size, moisture, melting resistance, the load difference, such as heavy metals and arsenic salts and microbial limit inspection were in line with the China pharmacopoeia2010version of an appendix I C granules under the relevant provisions of the general principles, and in its list of this product quality standard of text content. By high performance liquid chromatography (HPLC) method for the major effective components of indigotin, nitidine chloride and dictamnine content determination, and had carried on the system methodology, conformed to the requirements, and would be included in this product quality standard of text content. According to the determination results of several samples, formulated the main effective component content, tentative per bag (5.0g) of this product including indigotin in Pogostemon cablin (Blanco) Benth., shall not be less than773.08μg; nitidine chloride in Zanthoxylum nitidum (Roxb.) DC, shall not be less than846.84μg; dictamnine in Dictamnus dasycarpus Turcz., shall not be less than3301.67μg. Preliminary stability research, accelerated stability test results showed that, using aluminum-plastic laminated film bag samples used for the packaging materials during the investigation period the character, identification, examination and so on various examining indexes meet the product quality standard draft regulations. It suggested this product under the condition of high temperature and high humidity storage for six months, to ensure the quality stability. Long-term stability test results showed that the nine months of the period, the character, identification, examination and so on various examining indexes satisfy the requirements of this product quality standard. It suggested this product under the conditions of the save nine months, could ensure the stable quality. And the end of the test time was24month, the follow-up work was still in progress.5Study on chemical constituents of traditional Chinese medicine compound Fei kangmingAccording to the prescriptions of200times, took Baphicacanthus cusia (Nees) Bremek., Zanthoxylum nitidum (Roxb.) DC, Dictamnus dasycarpus Turcz., water reflux extraction, extracted after decompression enrichment and spray drying, and after the chloroform, ethyl acetate, n-butanol part of separation in turn, used silica gel column chromatography and recrystallization method to system of separation and purification, chemical composition and according to the physical and chemical constant determination and spectrometric (EI-MS or EIS-MS,’H-NMR,13C-NMR, Dept, etc.) the data and document control determine the structure of21monomer compounds, respectively were:nitidine chloride (1), chrysophanic acid (2), ursolic acid (3), indigotin (4), indirubin (5), stearic acid (6), fargesin (7), psoralen(8), luteolin(9), vanillin(10), palmitic acid(11), emodin-8-O-beta-D glucopyranoside(12), quercetin-3-rhamnoside(13), fraxinellone(14), obacunone(15), citric acid(16), chelidonine(17), adenosine(18), cytidine(19), baicah(20), benzoic acid(21). Of compounds3,7,10-13,16-17and19-21in the composition of traditional Chinese medicine compound Fei kangming for the first time found in3taste of Baphicacanthus cusia (Nees) Bremek., Zanthoxylum nitidum (Roxb.) DC., Dictamnus dasycarpus Turcz..To sum up, this topic through in vitro bacteriostasis experiment of10flavour Chinese medicine screening, to determine the best of traditional Chinese medicine compound, tentatively named compound Fei kangming, for MG infection had good treatment effect, used safety. Granule preparation technology was studied for the party, the craft was simple and reasonable, feasible and stable to meet the requirements of industrial production. This preparation was basic stability, high safety, stable and controllable quality standards. At the same time from the chloroform extract and ethyl acetate extract and n-butanol extract of the communist party of China to separate21compounds were identified, including11compounds were first in the composition of traditional Chinese medicine compound Fei kangming for the first time found in3taste of Baphicacanthus cusia (Nees) Bremek., Zanthoxylum nitidum (Roxb.) DC., Dictamnus dasycarpus Turcz.. This topic for the research and development and extensive application of new veterinary drug clinical animal mycoplasma infection disease prevention and control of traditional Chinese medicine provided a scientific basis.